SASDAQ4: 6:3 complex of GAD:CaM (Calmodulin + Glutamate decarboxylase, GAD)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDAQ4

試料

6:3 complex of GAD:CaM

• Calmodulin (protein), Homo sapiens
• Glutamate decarboxylase (protein), GAD, Petunia x hybrida

機能・相同性

分子機能:

glutamate decarboxylase / glutamate decarboxylase activity / : / : / : / : / : / positive regulation of protein autophosphorylation / L-glutamate catabolic process / negative regulation of peptidyl-threonine phosphorylation / : / type 3 metabotropic glutamate receptor binding / positive regulation of peptidyl-threonine phosphorylation / positive regulation of DNA binding / negative regulation of high voltage-gated calcium channel activity / response to corticosterone / negative regulation of ryanodine-sensitive calcium-release channel activity / negative regulation of calcium ion export across plasma membrane / regulation of cardiac muscle cell action potential / nitric-oxide synthase binding / regulation of synaptic vesicle exocytosis / regulation of cell communication by electrical coupling involved in cardiac conduction / adenylate cyclase binding / positive regulation of protein serine/threonine kinase activity / protein phosphatase activator activity / regulation of synaptic vesicle endocytosis / detection of calcium ion / regulation of cardiac muscle contraction / catalytic complex / regulation of cardiac muscle contraction by regulation of the release of sequestered calcium ion / enzyme regulator activity / activation of adenylate cyclase activity / phosphatidylinositol 3-kinase binding / positive regulation of nitric-oxide synthase activity / regulation of release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / titin binding / regulation of calcium-mediated signaling / voltage-gated potassium channel complex / calcium channel complex / substantia nigra development / regulation of heart rate / nitric-oxide synthase regulator activity / adenylate cyclase activator activity / response to amphetamine / sarcomere / protein serine/threonine kinase activator activity / regulation of cytokinesis / spindle microtubule / response to calcium ion / mitochondrial membrane / G2/M transition of mitotic cell cycle / disordered domain specific binding / spindle pole / calcium-dependent protein binding / pyridoxal phosphate binding / myelin sheath / synaptic vesicle membrane / growth cone / vesicle / transmembrane transporter binding / calmodulin binding / G protein-coupled receptor signaling pathway / protein domain specific binding / calcium ion binding / centrosome / protein kinase binding / protein-containing complex / nucleus / plasma membrane / cytoplasm / cytosol

ドメイン・相同性:

Glutamate decarboxylase / Pyridoxal phosphate-dependent decarboxylase / Pyridoxal-dependent decarboxylase conserved domain / EF-hand domain pair / Pyridoxal phosphate-dependent transferase, major domain / Pyridoxal phosphate-dependent transferase / EF-hand, calcium binding motif / EF-Hand 1, calcium-binding site / EF-hand calcium-binding domain. / EF-hand calcium-binding domain profile. / EF-hand domain / EF-hand domain pair

構成要素:

Calmodulin-3 / Glutamate decarboxylase

• 生物種: Homo sapiens (ヒト); Petunia x hybrida (ナス科)
• 引用: ジャーナル: J Mol Biol / 年: 2009; タイトル: A common structural basis for pH- and calmodulin-mediated regulation in plant glutamate decarboxylase.; 著者: Heinz Gut / Paola Dominici / Stefania Pilati / Alessandra Astegno / Maxim V Petoukhov / Dmitri I Svergun / Markus G Grütter / Guido Capitani / ; 要旨: Glutamate decarboxylase (Gad) catalyzes glutamate to gamma-aminobutyrate conversion. Plant Gad is a approximately 340 kDa hexamer, involved in development and stress response, and regulated by pH and binding of Ca(2+)/calmodulin (CaM) to the C-terminal domain. We determined the crystal structure of Arabidopsis thaliana Gad1 in its CaM-free state, obtained a low-resolution structure of the calmodulin-activated Gad complex by small-angle X-ray scattering and identified the crucial residues, in the C-terminal domain, for regulation by pH and CaM binding. CaM activates Gad1 in a unique way by relieving two C-terminal autoinhibition domains of adjacent active sites, forming a 393 kDa Gad1-CaM complex with an unusual 1:3 stoichiometry. The complex is loosely packed: thanks to the flexible linkers connecting the enzyme core with the six C-terminal regulatory domains, the CaM molecules retain considerable positional and orientational freedom with respect to Gad1. The complex thus represents a prototype for a novel CaM-target interaction mode. Thanks to its two levels of regulation, both targeting the C-terminal domain, Gad can respond flexibly to different kinds of cellular stress occurring at different pH values.

登録者

• Maxim Petoukhov (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #67: ; タイプ: mix / ソフトウェア: Bunch / ダミー原子の半径: 1.90 A / 対称性: P6/P1; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: 6:3 complex of GAD:CaM / Sample MW: 357.22 kDa / Entity id: 58 / 61
• バッファ: 名称: HEPES / 濃度: 50.00 mM / PK: 7 / pH: 7.5 ; コメント: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; 組成: KCl 50.000 mM

要素 #58

タイプ: protein / 記述: Calmodulin / 分子量: 16.84 / 分子数: 1 / 由来: Homo sapiens / 参照: UniProt: P62158
配列:

MADQLTEEQI AEFKEAFSLF DKDGDGTITT KELGTVMRSL GQNPTEAELQ DMINEVDADG NGTIDFPEFL TMMARKMKDT DSEEEIREAF RVFDKDGNGY ISAAELRHVM TNLGEKLTDE EVDEMIREAD IDGDGQVNYE EFVQMMTAK

要素 #61

名称: GAD / タイプ: protein / 記述: Glutamate decarboxylase / 分子量: 56.73 / 分子数: 6 / 由来: Petunia x hybrida / 参照: UniProt: Q07346
配列:

MVLSKTVSQS DVSIHSTFAS RYVRTSLPRF KMPDNSIPKE AAYQIINDEL MLDGNPRLNL ASFVTTWMEP ECDKLMMDSI NKNYVDMDEY PVTTELQNRC VNMIAHLFNA PLEDGETAVG VGTVGSSEAI MLAGLAFKRK WQNKMKAQGK PCDKPNIVTG ANVQVCWEKF ARYFEVELKE VKLSEGYYVM DPEKAVEMVD ENTICVAAIL GSTLNGEFED VKRLNDLLVE KNKETGWDTP IHVDAASGGF IAPFIYPELE WDFRLPLVKS INVSGHKYGL VYAGIGWVVW RNKDDLPDEL IFHINYLGAD QPTFTLNFSK GSSQVIAQYY QLIRLGYEGY KNVMENCQEN ASVLREGLEK TGRFNIISKE IGVPLVAFSL KDNRQHNEFE ISETLRRFGW IVPAYTMPPN AQHITVLRVV IREDFSRTLA ERLVRDIEKV LHELDTLPAR VNAKLAVAEE QAAANGSEVH KKTDSEVQLE MITAWKKFVE EKKKKTNRVC

実験情報

• ビーム: 設備名称: DORIS III X33 / 地域: Hamburg / 国: Germany / 線源: X-ray synchrotron
• 検出器: 名称: MAR 345 Image Plate

スキャン

タイトル: GAD:CaM complex / 測定日: 2004年11月9日 / 単位: 1/nm /

	Min	Max
Q	0.1557	4.483

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.6 / ポイント数: 285 /

	Min	Max
Q	0.1568	1.396
P(R) point	1	285
R	0	23.17

結果

I0 from Guinier: 87600 / Rg from Guinier: 5.85 nm / カーブのタイプ: single_conc /

	実験値	Porod
分子量	400 kDa	-
体積	-	630 nm3

/

	Min	Max
D	-	23
Guinier point	1	29